Stem Cell Isolation

Nozad H.Stem cell workshop

Stem cell Research center Tabriz medical university

In The Name of God

Isolation of ESCs1. Removing the outer trophectodermal cells by lysing using an antibody/complement reaction (“immunosurgery”).

2. Direct plating of the blastocyst without removal of the trophectoderm, but later requires the inner cell mass (ICM) cells away from the trophectodermal cells in vitro.3. In human routinly doing by IVF

Classic markers for evaluation of ESCs

1. Transcription markers: Nanog and Oct-4

2. Surface antigen stage–specific(SSEA3/4), tumor rejection antigen (TRA) 1-60, and TRA 1-81

Isolation of Mesenchaimal stem cell

Hematoppoietic stem cell isolation

1. It is 0.03% in bone marrow2. Because of very low it should first enriched or

deplet.Using MACS column for enrichment by pan surface marker: C-Kit in mice and CD34 in humanBind to a beads

Hematopoietic stem cell isolation

2. Depletion using dayna bead and magnet

Hematopoietic stem cell isolation

Sorting by FACS sorter

1. The enzymatic digestion is a critical step because it affects directly to the NSPc survival rate.

Neural stem cell isolation

2. The most common method for excluding the dead cells is passing the suspension through fire polished glass pipettes.

12-20 h.

Umblical Cord blood stem cell 1. Isolating mononucleur cell by ficoll paque gradiant

Umblical cord blood stem cell isolation

2. Enrichmet the CD34+ cells or CD133+ cells by magnetic beads Using MACS column

3. Sorting the specific population

Final point

The cell sorting by FACS is the best manner for Isolation of Stem cells because of high purity

Thank you very much for your attention Good Luck